微囊化血管内皮细胞生长因子基因修饰NIH3T3细胞的制备及体外培养

Preparation and culture of NIH3T3 cells harboring microencapsulated VEGF gene

目的:制备微囊化血管内皮细胞生长因子(VEGF)基因修饰NIH3T3细胞,并研究微囊化技术对VEGF基因修饰NIH3T3细胞增殖及其分泌VEGF功能的影响。方法:应用海藻酸钠-氯化钡技术制备微囊化VEGF基因修饰NIH3T3细胞。并与未微囊化VEGF基因修饰NIH3T3细胞对照培养,在倒置相差显微镜下观察微囊及细胞形态,用MTT法和PI染色流式细胞术检测细胞的增殖及活性情况。每48 h收集微囊化及未微囊化基因修饰NIH3T3细胞培养上清,-20℃保存.ELISA法检测培养上清VEGF的含量。结果:微囊形态较圆整,其内细胞生长良好;两组细胞增殖与活性及培养上清中VEGF含量无统计学差异。结论:微囊化并不影响基因修饰细胞的生长代谢功能,与对照组比较.在体外培养时生物学特性无明显差异,为研究微囊化基因修饰细胞体内移植奠定了基础。

Objective：To prepare NIH3T3 cells harboring microencapsulated VEGF gene and investigate the proliferation, activity and metabolic function of the modified cells. Methods： Microencapsulated VEGF modified NIH3T3 cells were prepared through an alginate-BaCl2 process. Morphological appearance of the microencapsulation and the cell morphology were observed under inverted phase microscope; untreated NIH3T3 cells served as control. The concentrations of VEGF in the culture superna- rant （collected every 48 hours） were measured by ELISA; the proliferation and vitality of the cells were examined by MTT as- say and flow cytometry with PI staining. Results： The microcapsules were round in shape and the cells grew well. There was no significant difference in the concentrations of VEGF,MTT values and vitalities of cells between the 2 groups. Conclusion： The growth and metabolic functions of NIH3T3 cells are not influenced by microencapsulated NIH3T3 cells harboring VEGF gene. The bio-properties of modified cells are similar to those of the control cells,which lays a foundation for transplantation of microencapsulated VEGF modified NIH3T3 cells in vivo.